A dedicated microarray for in-depth analysis of pre-mRNA splicing events: application to the study of genes involved in the response to targeted anticancer therapies

Table 3 Quantitative RT-PCR validation in SRSF2-over-expressing H358 lung adenocarcinoma cells

SRSF2 Condition vs. Control condition
Gene	Calculation	Transcript	Expression	Observed transcript regulation	Expected transcript regulation
HER1/EGFR	Relative expression	Last exon = e17	n/a	Not expressed	No expression
		Last exon = e18	0.91	Not regulated	Over-expression
		Last exon > e20	0.42	Under-expressed	Under-expression
AKT3	Fold-change	e7+ e8- vs. e7+ e8+	1.46	Over-expression of e7+ e8-	Over-expression of exon 7 and under-expression of exon 8
		e7- e8+ vs. e7+ e8+	-1.19	No regulation of e7- e8+
		e7- e8- vs. e7+ e8+	n/a	No expression of e7- e8-
HIF1A	Fold-change	e9+ e10- vs. e9+ e10+	1.91	Over-expression of e9+ e10-	Under-expression of exon 10
		e9- e10+ vs. e9+ e10+	-1.52	Under-expression of e9- e10+	Under-expression of exon 10
		e9- e10- vs. e9+ e10+	1.82	Over-expression of e9- e10-	Under-expression of exons 9 and 10
VEGFA	Fold-change	Last exon = e4 vs. last exon > e5	18.93	Over-expression of "last exon = e4"	Over-expression of exon 4
VEGFA	Fold-change	Alternative vs. constitutive donor e6	14.46	Over-expression of "alternative donor e6"	Over-expression of alternative donor

The regulation of the 10 selected deregulated custom probe sets was analyzed by quantitative RT-PCR in SRSF2-over-expressing lung cancer cells in comparison to control cells. Relative mRNA levels were normalized to that of beta-2-microglobulin or a fold-change was calculated comparing to a reference event. The cut-off value was equal to 1.40. n/a: not available.

ISSN: 1476-4598